灯盏花素上调核因子相关因子-2通路抑制液压冲击脑损伤炎性反应

Breviscapine alleviates neuroinflammation via modulating nuclear factor erythroid 2-related factor 2-ARE signaling pathway in rats following fluid percussion brain injury

目的:探讨灯盏花素上调核因子相关因子-2(Nrf2)通路抑制大鼠液压冲击脑损伤后脑组织炎性损伤的具体机制。方法:成年雄性SD大鼠36只(河北医科大学实验动物中心),应用Dixon法制作颅脑损伤模型,造模后24 h用干湿重法、酶联免疫吸附试验(ELISA)法和蛋白质印迹法(Western blot)分别测定脑组织含水量、炎性因子白细胞介素(IL)-1β、IL-6和肿瘤坏死因子-α(TNF-α)以及胞核Nrf2、血红素氧化酶1(HO-1)、磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶1(NQO-1)蛋白表达。应用SPSS 22.0统计软件分析,计量资料用均值±标准差( Mean± SD)表示,组间比较差异性分析采用Student’s t检验。 结果:与模型组[创伤性脑损伤(TBI)组]比较,灯盏花素组(Bre组)造模后24 h脑组织含水量[(81.82±2.64)%比(83.04±1.89)%, t=2.335, P<0.05]和改良大鼠神经功能缺损评分(mNSS)[(7.98±1.68)分比(9.61±1.74)分, t=5.271, P<0.05]均明显降低;ELISA结果显示Bre组炎性因子IL-1β[(834.19±95.68) ng/L比(1213.35±167.72) ng/L, t=6.802, P<0.05]、IL-6[(627.82±86.54) ng/L比(1336.72±198.25) ng/L, t=9.643, P<0.05]和TNF-α[(586.64±76.98) ng/L比(1416.67±136.85) ng/L, t=10.312, P<0.05]表达明显降低;Western blot结果显示Bre组胞核Nrf2[(1.52±0.34)比(0.49±0.09), t=5.139, P<0.05]、HO-1[(0.87±0.21)比(0.49±0.09), t=5.762, P<0.05]和NQO-1[(1.46±0.36)比(0.99±0.19), t=4.998, P<0.05]蛋白表达明显上调。 结论:灯盏花素通过激活Nrf2-抗氧化元件(ARE)信号系统,上调下游抗氧化蛋白HO-1和NQO-1,抑制炎性损伤。

Objective To investigate the mechanism of Breviscapine in alleviating neuroinflammation via modulating nuclear factor erythroid 2-related factor 2(Nrf2)-ARE signaling pathway in rats following fluid percussion brain injury.Methods The experimental models were established in 36 rats according to Dixon’s method.Brain water content,inflammatory cytokines[interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)],the expression levels of nucleus Nrf2,heme oxygenase-1(HO-1)and NQO-1 proteins were measured by dry-wet measure,enzyme linked immunosorbent assay(ELISA)and Western blotting.Results As compared with traumatic brain injury(TBI)group,brain water content[(81.82±2.64)%vs.(83.04±1.89)%,t=2.335,P<0.05]and modified neurological severity scores(mNSS)[(7.98±1.68)vs.(9.61±1.74),t=5.271,P<0.05]at 24 hwere significantly decreased,the expression levels of inflammatory cytokines IL-1β[(834.19±95.68)vs.(1213.35±167.72)ng/L,t=6.802,P<0.05],IL-6[(627.82±86.54)vs.(1336.72±198.25)ng/L,t=9.643,P<0.05]and TNF-α[(586.64±76.98)vs.(1416.67±136.85)ng/L,t=10.312,P<0.05]significantly down-regulated,and the protein expression levels of nucleus Nrf2[(1.52±0.34)vs.(0.49±0.09),t=5.139,P<0.05],HO-1[(0.87±0.21)vs.(0.49±0.09),t=5.762,P<0.05]and NQO-1[(1.46±0.36)vs.(0.99±0.19),t=4.998,P<0.05]were up-regulated in Breviscapine group.Conclusion Breviscapine inhibits neuroinflammatory injury via modulating Nrf2-antioxidant response element pathway including the expression of HO-1 and NQO-1 proteins in rats following fluid percussion brain injury.